.beta.-sarcoglycan nucleic acid sequence, and nucleic acid probes
Disclosed herein is a substantially pure nucleic acid sequence encoding a mammalian 43 kDa non-dystrophin component (.beta.-sarcoglycan) of the dystrophin-glycoprotein complex. Also disclosed are immunogenic peptides which, when used to immunize a mammal, stimulate the production of antibodies which bind specifically to the .beta.-sarcoglycan. Mutations in the .beta.-sarcoglycan gene which are associated with autosomal recessive limb-girdle muscular dystrophy are also disclosed. The identification of such mutations enables the design of nucleic acid probes which hybridize specifically to a mutant form of .beta.-sarcoglycan, or the complement thereof, but not to the DNA of the wild-type form of the gene (or the complement thereof), under stringent hybridization conditions. Such probes are useful, for example, in connection with the diagnosis of autosomal recessive limb-girdle muscular dystrophy. In addition, the identification of such mutations enables the diagnosis of autosomal recessive limb-girdle muscular dystrophy through the use of direct DNA sequencing techniques.
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Claims
1. A substantially pure nucleic acid sequence encoding a mammalian 43 kDa non-dystrophin component of the dystrophin-glycoprotein complex which is characterized by the ability to hybridize to the DNA sequence of SEQ ID NO:1, or the fully complementary sequence thereof, under stringent hybridization conditions.
2. A substantially pure nucleic acid sequence of claim 1 which is of human origin.
3. A DNA expression construct comprising, in expressible form, a substantially pure deoxyribonucleic acid sequence encoding a mammalian 43 kDa non-dystrophin component of the dystrophin-glycoprotein complex which is characterized by the ability to hybridize to the DNA sequence of SEQ ID NO:1, or the fully complementary sequence thereof, under stringent hybridization conditions.
4. A DNA expression construct of claim 3 wherein the substantially pure nucleic acid is of human origin.
5. A prokaryotic cell transformed with a DNA expression construct comprising, in expressible form, a substantially pure deoxyribonucleic acid sequence encoding a mammalian 43 kDa non-dystrophin component of the dystrophin-glycoprotein complex which is characterized by the ability to hybridize to the DNA sequence of SEQ ID NO:1, or the fully complementary sequence thereof, under stringent hybridization conditions.
6. A prokaryotic cell of claim 5 wherein the substantially pure deoxyribonucleic acid sequence is of human origin.
7. A eukaryotic cell transformed with a DNA expression construct comprising, in expressible form, a substantially pure deoxyribonucleic acid sequence encoding a mammalian 43 kDa non-dystrophin component of the dystrophin-glycoprotein complex which is characterized by the ability to hybridize to the DNA sequence of SEQ ID NO:1, or the fully complementary sequence thereof, under stringent hybridization conditions.
8. A eukaryotic cell of claim 7 wherein the substantially pure deoxyribonucleic acid sequence is of human origin.
9. A substantially pure nucleic acid molecule, or the fully complementary sequence complement thereof, the substantially pure nucleic acid molecule encoding the amino acid sequence shown in SEQ ID NO:2.
10. A substantially pure nucleic acid molecule of claim 9 which is of human origin.
11. A DNA expression construct comprising, in expressible form, a substantially pure nucleic acid molecule encoding the amino acid sequence shown in SEQ ID NO:2.
12. A DNA expression construct of claim 11 wherein the substantially pure nucleic acid molecule is of human origin.
13. A prokaryotic cell transformed with a DNA expression construct comprising, in expressible form, a substantially pure nucleic acid molecule encoding the amino acid sequence shown in SEQ ID NO:2.
14. A eukaryotic cell transformed with a DNA expression construct comprising, in expressible form, a substantially pure nucleic acid molecule encoding the amino acid sequence shown in SEQ ID NO:2.
15. A nucleic acid probe of at least 20 nucleotides which hybridizes specifically to a mutant form of.beta.-sarcoglycan, or the fully complementary sequence thereof, but not to the DNA of SEQ ID NO:1 or the fully complementary sequence thereof, under stringent hybridization conditions, the mutant form differing from the wild-type form of.beta.-sarcoglycan in a change from threonine to arginine at codon 151 shown in SEQ ID NO:1.
16. A nucleic acid probe of at least 20 nucleotides which hybridizes specifically to a mutant form of.beta.-sarcoglycan, or the fully complementary sequence thereof, but not to the DNA of SEQ ID NO:1 or the fully complementary sequence thereof, under stringent hybridization conditions, the mutant form differing from the wild-type form of.beta.-sarcoglycan in a change from serine to phenylalanine at codon 114 shown in SEQ ID NO:1.
17. A nucleic acid probe of at least 20 nucleotides which hybridizes specifically to a mutant form of.beta.-sarcoglycan, or the fully complementary sequence thereof, but not to the DNA of SEQ ID NO:1 or the fully complementary sequence thereof, under stringent hybridization conditions, the mutant form differing from the wild-type form of.beta.-sarcoglycan in a change from isoleucine to phenylalanine at codon 119 shown in SEQ ID NO:1.
- Speer et al., Am. J. Hum. Genet. 50: 1211 (1992). Bashir et al. Hum. Mol. Genet. 3: 455 (1994). Ben Othmane et al., Nature Genet. 2: 315 (1992). Azibi et al., Hum. Mol. Genet. 2: 1423 (1993). Beckmann et al., C.R. Acad. Sci. Paris 312: 141 (1991). Roberds et al., Cell 78: 625 (1994). Richard et al., Cell 81: 27 (1995). New England Biolab Catalog, Beverly, MA, USA, p. 56 and p. 61 1986. Sigma Chemical Company Catalog, p. 1639 1990.
Type: Grant
Filed: Oct 24, 1995
Date of Patent: Sep 30, 1997
Assignee: University of Iowa Research Foundation (Iowa City, IA)
Inventors: Kevin P. Campbell (Iowa City, IA), Leland Lim (Iowa City, IA), Franck Duclos (Iowa City, IA), Yoshihide Sunada (Iowa City, IA), Jacques S. Beckmann (Charenton-le-Pont), Odile Broux (L'Hay-les-Roses), Fernando M. S. Tome (Paris), Michel Fardeau (Sceaux), Charles E. Jackson (Grosse Pointe, MI)
Primary Examiner: Ardin H. Marschel
Assistant Examiner: Jezia Riley
Attorney: Kevin M. Farrell
Application Number: 8/547,182
International Classification: C07H 1900; C07H 2102; C07H 2104; C12Q 168;
